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Microbial enhanced oil recovery (EOR) has become the focus of oilfield research due to its low cost, environmental friendliness and sustainability. The degradation and EOR capacity of A. borkumensis through the production of bio-enzyme and bio-surfactant were first investigated in this study. The total protein concentration, acetylcholinesterase, esterase, lipase, alkane hydroxylase activity, surface tension, and emulsification index (EI) were determined at different culture times. The bio-surfactant was identified as glycolipid compound, and the yield was 2.6 ± 0.2 g/L. The nC12 and nC13 of crude oil were completely degraded, and more than 40.0 % of nC14-nC24 was degraded by by A. borkumensis. The results of the microscopic etching model displacement and core flooding experiments showed that emulsification was the main mechanism of EOR. A. borkumensis enhanced the recovery rate by 20.2 %. This study offers novel insights for the development of environmentally friendly and efficient oil fields.

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The current work reports the gum arabic-mediated greener synthesis of MoS2 nanoparticles (NPs) and its utilization for the solar light-assisted degradation of methylene blue. Furthermore, the safety analyses were performed on human-beneficial gut bacterium, L. delbrueckii, and human blood cells to confirm the biocompatibility of NPs synthesized. Antioxidant and antimicrobial activities were done to explore their usefulness for biological applications. Sonication and microwave treatment were used to obtain spherical 10-12 nm MoS2 NPs as characterized using high-resolution transmission electron microscopy. FT-IR characterization revealed the occurrence of gum arabic on the NPs surface. The MoS2 NPs exhibited ~ 98% MB degradation within 8 h under direct sunlight exposure. Moreover, the reusability studies have also been evaluated and free radical trapping experiments indicated that superoxide (•O2-) is the dominant active species of the reaction system. Furthermore, 98.89% MB degradation efficiency was observed within 150 min in the case of real textile industry MB effluent samples. Untreated MB inhibited the growth of L. delbrueckii on MRS agar plates, while growth was observed in the case of MoS2 NPs-treated MB samples indicating safety of current MB degradation approach. MoS2 NPs inhibited the growth of E. coli MTCC1698 and S. aureus MTCC 3160 with 26 mm and 21 mm zone of inhibition, respectively. Furthermore, MoS2 NPs have shown antioxidant properties, resulting in 82.3 ± 0.43% of DPPH scavenging activity which was comparable to ascorbic acid (81.6 ± 0.6%), a standard antioxidant molecule. The NPs have not shown any hemolytic activity at 0.0625 and 0.125 mg/mL doses to human blood proving their biocompatible nature. Gum arabic-synthesized biocompatible MoS2 NPs have good potential to treat MB released as waste from the textile industry and other biological applications.

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Landfill leachate-contaminated soil is widespread all over the world. In order to study the removal of mixed contaminants from landfill leachate-contaminated soil by flushing with bio-surfactant, soil column test was conducted to select an optimum concentration of bio-surfactant saponin (SAP) at first. Then, the removal efficiencies of organic contaminants, ammonia nitrogen, and heavy metals from landfill leachate-contaminated soil by flushing with SAP were studied. At last, the toxicity of contaminated soil before and after flushing was estimated by sequential extraction of heavy metals and plant growth test. The test results showed that the SAP solution with the concentration of 2.5 CMC could effectively remove the mixed contaminants from soil and would not introduce excessive pollutants of SAP in soil. Specifically, the removal efficiencies of organic contaminant and ammonia nitrogen were 47.01% and 90.42%, respectively. And the removal efficiencies of Cu, Zn, and Cd were 29.42%, 22.55%, and 17.68%, respectively. During flushing, hydrophobic organic compounds as well as physisorption and ion-exchange ammonia nitrogen in soil were removed by the solubilization effect of SAP, and heavy metals were removed by the chelation of SAP. After flushing with SAP, the reduced partition index (IR) value of Cu and Cd increased, and the mobility index (MF) value of Cu decreased. In addition, flushing with SAP reduced the plant toxicity of contaminated soil, and the residual SAP in soil promoted the plant growth. Therefore, flushing with SAP offered great potentials in remediating the landfill leachate-contaminated soil.

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Bio surfactants are natural surfactants that induce emulsification, displacement, increased solubility, and mobility of hydrophobic organic compounds. In this study, the gene expression of biosurfactant production genes by Pseudomonas aeruginosa in the presence of sodium dodecyl sulfate coated iron nanostructure (Fe/SDS) were evaluated. Emulsification Index and Surface Tension reduction test to check stability and emulsification the rhamnolipid were done. Purification was evaluated using thin layer chromatography (TLC) and expression of rhlA, mvfR, lasR, rhlR genes was determined using q-PCR technique. Binding of nanoparticles to bio surfactants was confirmed by TEM. The best emulsification index, was by the sample that exposed to 1 mg/L Fe/SDS nanoparticles for 2 days. Rhamnolipid produced in the presence of nanoparticles had an acceptable ability to reduce surface tension. The Rf (retention factor) value obtained was 0.63 by chromatography. q-PCR results showed that the expression of rhlA, mvfR, lasR, rhlR genes was significantly increased in Fe/SDS treated cells, which indicates the significant positive effect (P < 0.05) of nanoparticles on biosurfactant production of treated cells. While, SDS and Fe alone were not affected significantly (P > 0.05) on the expression of these genes. Our findings indicated the importance of nanoparticles in increasing the expression of genes involved in the bio surfactant production pathway of Pseudomonas aeruginosa.

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Soil salinity is one of the abiotic constraints that imbalance nutrient acquisition, hampers plant growth, and leads to potential loss in agricultural productivity. Salt-tolerant plant growth-promoting rhizobacteria (PGPR) can alleviate the adverse impacts of salt stress by mediating molecular, biochemical, and physiological status. In the present study, the bacterium Bacillus mycoides PM35 showed resistance up to 3 M NaCl stress and exhibited plant growth-promoting features. Under salinity stress, the halo-tolerant bacterium B. mycoides PM35 showed significant plant growth-promoting traits, such as the production of indole acetic acid, siderophore, ACC deaminase, and exopolysaccharides. Inoculation of B. mycoides PM35 alleviated salt stress in plants and enhanced shoot and root length under salinity stress (0, 300, 600, and 900 mM). The B. mycoides PM35 alleviated salinity stress by enhancing the photosynthetic pigments, carotenoids, radical scavenging capacity, soluble sugars, and protein content in inoculated maize plants compared to non-inoculated plants. In addition, B. mycoides PM35 significantly boosted antioxidant activities, relative water content, flavonoid, phenolic content, and osmolytes while reducing electrolyte leakage, H2O2, and MDA in maize compared to control plants. Genes conferring abiotic stress tolerance (CzcD, sfp, and srfAA genes) were amplified in B. mycoides PM35. Moreover, all reactions are accompanied by the upregulation of stress-related genes (APX and SOD). Our study reveals that B. mycoides PM35 is capable of promoting plant growth and increasing agricultural productivity.

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Mannosylerythritol lipid-B (MEL-B), which comprises ester-bonded hydrophilic ME and hydrophobic fatty acids, is a bio-surfactant with various unique properties, including antimicrobial activity against most gram-positive bacteria. The gram-positive Staphylococcus aureus is a causative pathogen of dairy cattle mastitis, which results in considerable economic loss in the dairy industry. Here, we demonstrate the efficacy of MEL-B as a disinfectant against bovine-derived S. aureus and elucidate a mechanism of action of MEL-B in the inhibition of bacterial growth. The growth of bovine mastitis causative S. aureus BM1006 was inhibited when cultured with MEL-B above 10 ppm. The activity of MEL-B required fatty acids (i.e., caprylic and myristoleic acids) as ME, the component of MEL-B lacking fatty acids, did not inhibit the growth of S. aureus even at high concentrations. Importantly, ME-bound fatty acids effectively inhibited the growth of S. aureus when compared with free fatty acids. Specifically, the concentrations of ME-bound fatty acids and free caprylic and myristoleic acids required to inhibit the growth of S. aureus were 10, 1442, and 226 ppm, respectively. The involvement of ME in the antimicrobial activity of MEL-B was confirmed by digestion of MEL-B with alkali, which dissociated ME and fatty acids. These results indicated that a mechanism of action of MEL-B in inhibiting the growth of S. aureus could be explained by the effective transporting of antimicrobial fatty acids to the bacterial surface via hydrophilic ME.

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Biohydrogen production from marine macroalgal biomass by advanced pre-treatment strategies is considered a clean energy technology. The present study focuses on investigating the effects of sonication pre-treatment (SP) and saponin coupled sonic pre-treatment (SSP) on Ulva fasciata for enhancing the production of biohydrogen. The SP and SSP were optimized to improve the hydrolysis process during digestion. The optimized time and sonication power were found respectively as 30 min and 200 W. A high concentration of biopolymer release was noticed in SSP than SP at optimized conditions. The surfactant dosage in SSP was optimized at 0.0036 g/g TS. The effect of SSP process was assessed by estimation of COD (Chemical Oxygen Demand) and SCOD (Soluble Chemical Oxygen Demand) release. The study revealed that, at a specific energy of 36,000 KJ/Kg TS, the SCOD release was higher in SSP (1900 mg/L) than SP (1050 mg/L). The SSP process could improve the COD solubilization to 15 % more than the SP. Carbohydrate and protein release are also more in SSP than SP. The use of biosurfactants significantly reduced the energy utilization in the hydrolysis process. The SSP pre-treated Ulva fasciata biomass has yielded a higher biohydrogen of 91.7 mL/g COD which is higher compared to SP (40.5 mL/g COD) and Control (9 mL/g COD).

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This work aims to produce bio-surfactant using a brewery waste (trub) as a strategy to reduce production costs related to the substrate, as well as to provide an eco-friendly destination for this residue. Trub is obtained during the boiling of the wort, being mainly composed of proteins and reducing sugars. To evaluate important process parameters on bio-surfactant production, a full factorial design (24) was elaborated, having agitation rate and concentrations of trub, yeast extract, and peptone as independent variables. The highest bio-surfactant concentration achieved was 100.76 mg L-1, where FTIR and Maldi-ToF-MS confirmed functional groups characteristic of peptides and isomers of surfactin in the bio-surfactant extract. Trub, agitation and yeast extract showed statistically significant effects on the response variable (surface tension), where an increase in the agitation rate and in the concentration of yeast extract demonstrated a positive impact on the production of bio-surfactant.

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Diesel removal of contaminated soil by washing/flushing was enhanced with micro-bubbles and selected surfactants based on their solubilization properties and decontamination capacities. The influencing factors were studied to aim for increasing washing/flushing efficacy. The mixture solution of saponin and cyclodextrin increased the removal efficiency significantly compared to the single-agent solution flushing with an increasing range of 20%-31%. Meanwhile, micro-bubble enhancement increased over 20% of the diesel removal for the sandy soil flushing. As the flushing process may cause soil eroded, the TDS and soil solute in flushing solution were measured to evaluate the circulation time. The 90 min flushing time ensured the cleaning goal and reserved the soil solute by circulation flushing. The soil solute, especially the electron acceptor (NO3-) , was remained in the soil, which was highly demanded for residual diesel biodegradation of loam soil. It is concluded that mixed agents, circulation of flushing solution, and micro-bubbles increased the diesel removal, and the circulation flushing could be very promising in practical applications.

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Methyl-mannosylerythritol lipid (MEL), a new sugar esterified lipid synthesized by Pseudozyma aphidis, was assessed for its functionality in modulating rumen fermentation and microbiota toward more propionate and less methane production. A pure culture study using rumen representatives showed that MEL selectively inhibited the growth of most Gram-positive bacteria including Streptococcus bovis, ruminococci, and Fibrobacter succinogenes, but not Gram-negative bacteria such as Megasphaera elsdenii, Succinivibrio dextrinosolvens, and Selenomonas ruminantium. A batch culture study revealed that MEL significantly decreased methane production in a dose-dependent manner with accumulation of hydrogen, while propionate production was enhanced. A continuous culture (Rusitec) study confirmed all of these changes. A feeding study revealed that sheep fed a MEL diet showed an increased proportion of propionate, while proportions of acetate and butyrate were decreased without affecting total VFA level. These changes disappeared after cessation of MEL feeding. Based on these results, dietary application of MEL can favorably modify rumen fermentation in terms of the efficiency of dietary energy utilization.

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The aim of this study is to fabricate a highly porous scaffold based on gelatin/sodium alginate/45S5 bioglass with improved mechanical strength. Glycidoxypropyltrimethoxysilane (GPTMS) is used as a cross-linker and for the first time a nonionic bio-surfactant (Tween 80) is introduced to the composition to evaluate whether the essential properties of a suitable scaffold for bone substitution can be reached or not. The composite scaffolds are prepared through freeze-drying of suspension containing various ratios of gelatin/sodium alginate/45S5 bioglass. Characterization of fabricated scaffolds is carried out. SEM micrographs reveal that all samples are highly porous however incorporation of 1% v/v tween 80 results in well-shaped pores, with sizes ranging between 100 and 200 µm which is appropriate for tissue regeneration. Compressive strength of foamed scaffolds in contact with body solution has been enhanced from 0.37 to 1.41 MPa due to addition of tween 80. Foamed scaffold reinforced with tween 80 maintained its structural stability within 7 days immersion in simulated body fluid (SBF) despite the absorption of water 15 times its weight. Moreover, in vitro calcium phosphate precipitation is well observed on the surface of scaffolds.

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Optimising detergency at lower temperatures is of increasing interest due to environmental and economic factors, and requires a greater understanding of the effects of temperature on the adsorption of surfactant mixtures at interfaces. The adsorption properties of surfactant mixtures and biosurfactant/surfactant mixtures have been studied at room temperatures and at temperatures below ambient using surface tension and neutron reflectivity measurements. For the ternary surfactant mixture of octaethylene monododecyl ether, C12E8, sodium dodecyl 6-benzene sulfonate, LAS, and sodium dioxyethylene glycol monododecyl sulfate, SLES, the surface tension at the air-water interface increases with decreasing temperature. In contrast, there is a notable reduction in the increase in the surface tension with a decrease in temperature from 25 °C to 10 °C for the 5 component rhamnolipid/surfactant mixture of the mono-rhamnose, R1, and di-rhamnose, R2, with C12E8/LAS/SLES. The associated neutron reflectivity data for the ternary C12E8/LAS/SLES mixture and the significant observation is that the 3, 4, and 5-component mixtures containing rhamnolipids in conjunction with the other surfactants show changes in composition and adsorbed amounts of the individual components which are close to the experimental error. However the significant observation is that the neutron reflectivity data indicate that the improved surface tension tolerance at lower temperatures is associated with the dominance of the rhamnolipid adsorption in such mixtures. Hence the introduction of the rhamnolipids provides a tolerance to the adverse effects associated with reduced temperatures, and a potential for improved detergency at relatively low temperatures.

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In this study, a trehalose lipid was added to a Rhodococcus pyridinivorans-inoculated MFC to improve the power output by enhancing electron transfer. Upon trehalose lipid additions of different concentrate from 0 to 20 mg/L, the maximum power density increased from 54.7 mW/m2 to 324.4 mW/m2 (5.93 times) while the corresponding current density was 3.66 times increased from 0.35 A/m2 to 1.28 A/m2. Cyclic voltammetry analysis revealed that the addition of trehalose lipid increased the electron transfer performance, while electrochemical impedance spectroscopy results proved a decrease in internal resistance. It was demonstrated that adding bio-surfactant in MFC was a novel way to enhance power output performance.

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Aggregation behavior of bio-surfactants (BS) sodium cholate (NaC) and sodium deoxycholate (NaDC) within aqueous solution of ionic liquid (IL) 1-ethyl-3-methylimidazolium bromide [Emim][Br] has been investigated using surface tension, conductivity, steady state fluorescence, FT-IR and dynamic light scattering (DLS) techniques. Various interfacial and thermodynamic parameters are determined in the presence of different wt% of IL [Emim][Br]. Information regarding the local microenvironment and size of the aggregates is obtained from fluorescence and DLS, respectively. FT-IR spectral response is used to reveal the interactions taking place within aqueous NaC/NaDC micellar solutions. It is noteworthy to mention that increasing wt% of [Emim][Br] results in an increase in the spontaneity of micelle formation and the hydrophilic IL shows more affinity for NaC as compared to NaDC. Further, the micellar solutions of BS-[Emim][Br] are utilized for studying the aggregation of antidepressants drug promazine hydrochloride (pH). UV-vis spectroscopic investigation reveals interesting outcomes and the results show changes in spectral absorbance of PH drug on the addition of micellar solution (BS-[Emim][Br]). Highest binding affinity and most promising activity are shown for NaC as compared to NaDC.

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Dietzia sp. A14101 belonging to the genus Dietzia (Gram-positive bacteria, Actinomycetes, high G+C content of DNA) was isolated from an oil reservoir model column inoculated with oil-field bacteria (Bødtker et al., 2009). Low interfacial tension (IFT) values were obtained by studying intact strain cells grown on water-immiscible hydrocarbons (HC) (Kowalewski et al. (2004), Kowalewski et al. (2005). Further investigations showed that the adaptation mechanism of Dietzia sp. A14101 to toxic water-immiscible HC involved changes both on the level of fatty acids content and in the physical properties of the cellular surface (development of the negative cellular surface charge and an increased in hydrophobicity) (Hvidsten et al., 2015b). However, these changes could not explain the low IFT values observed in earlier studies of this strain. Generally, low IFT imply a production of surface active compounds of low MW that are lipids by structure (Rosenberg and Ron, 1999). In this paper, it is shown that Dietzia sp. A14101 produces a range of glycolipids on all substrates. The amount of trehalose-containing lipids increases when the strain is grown on hydrocarbons. The production peak seems to coincide with the exponential growth phase, and such increased glycolipid synthesis continues throughout the stationary phase. The results indicate that only low amounts of the hydrocarbon substrate is incorporated directly into the glycolipids produced. Most of the hydrocarbon substrate seems to be employed for the biosynthesis of the neutral lipids and higher amounts of biomass were generated on HC substrates compared to incubations on non-HC substrates. The lipid content of the cell was determined as the total lipid extract (TLE), and was further fractionated (SPE). The hydrophobic and hydrophilic moieties of the isolated surface active compounds were determined (GC-MS, TLC, DART, LC-MS). The changes in the lipid content during the culture development were monitored by 1D and 2D TLC, emulsification and oil-spreading tests.

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Dietzia sp. A14101 isolated from an oil reservoir model column was found to induce a strong decrease of the interfacial tension (IFT) in hydrocarbon-water mixtures in the presence of the intact bacterial cells (Kowalewski et al., 2005). The strain was shown to be able to degrade a wide range of hydrocarbon substrates (Bødtker et al., 2009). Further studies showed that the surface-active compounds tentatively identified as glycolipids were produced by Dietzia sp. A14101 on non- and water-immiscible -hydrocarbon substrates, Part I (Hvidsten et al., 2017). The results suggested that biosurfactant (BS) was a mixture of several isomers. The study presented here is aimed to investigate whether BS are secreted into the aqueous medium, and if so, then at which phase of the culture growth and in which amounts - the dynamics of the BS release in incubations on water-immiscible hydrocarbons. Two methods of BS extraction from the medium were attempted and compared: a liquid-liquid extraction (LLE) and precipitation by acid. For qualitative and semi-quantitative assessment, gas chromatography-mass spectrometry (GC/MS), thin-layer chromatography (TLC), liquid chromatography-mass spectrometry (LC-MS), surface tension measurements (SFT), emulsification (E24) and oil-spreading tests were employed. The results indicated that BS only partially were secreted into the medium. Detectable amounts of glycolipids in media were first identified during the exponential growth phase. However, only a slight decrease of SFT was observed in the cell-free medium. The emulsification index values of the sampled material were lower than those reported for related strains. The results suggested that most of the BS produced by Dietzia sp. A14101 remains cell-bound during the culture development in a batch mode and only a narrow range of the BS isomers can be detected in small amounts in media.

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Halomonas spp. have been studied as a low cost production host for producing bulk materials such as polyhydroxyalkanoates (PHA) bioplastics, since they are able to grow at high pH and high NaCl concentration under unsterile and continuous conditions without microbial contamination. In this paper, Halomonas strain TD is used as a host to produce a protein named PHA phasin or PhaP which has a potential to be developed into a bio-surfactant. Four Halomonas TD expression strains are constructed based on a strong T7-family expression system. Of these, the strain with phaC deletion and chromosomal expression system resulted in the highest production of PhaP in soluble form, reaching 19% of total cellular soluble proteins and with a yield of 1.86 g/L in an open fed-batch fermentation process. A simple "heat lysis and salt precipitation" method is applied to allow rapid PhaP purification from a mixture of cellular proteins with a PhaP recovery rate of 63%. It clearly demonstrated that Halomonas TD could be used for high yield expression of a bio-surfactant protein PhaP for industrial application in an economical way.

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Sugar fatty acid esters are bio-surfactants known for their non-toxic, non-ionic, and high biodegradability . With great emulsifying and conditioning effects, sugar fatty acids are widely used in the food, pharmaceutical, and cosmetic industries. Biosynthesis of sugar fatty acid esters has attracted growing attention in recent decades. In this study, the enzymatic synthesis of sugar fatty acid esters in ionic liquids was developed, optimized, and scaled up. Reaction parameters affecting the conversion yield of lipase-catalyzed synthesis of glucose laurate from glucose and vinyl laurate (i.e. temperature, vinyl laurate/glucose molar ratio, and enzyme loads) were optimized by response surface methodology (RSM). In addition, production was scaled up to 2.5 L, and recycling of enzyme and ionic liquids was investigated. The results showed that under optimal reaction conditions (66.86 °C, vinyl laurate/glucose molar ratio of 7.63, enzyme load of 73.33 g/L), an experimental conversion yield of 96.4% was obtained which is close to the optimal value predicted by RSM (97.16%). A similar conversion yield was maintained when the reaction was carried out at 2.5 L. Moreover, the enzymes and ionic liquids could be recycled and reused effectively for up to 10 cycles. The results indicate the feasibility of ionic liquids as novel solvents for the biosynthesis of sugar fatty acid esters.

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